Skip to main content

Production of SV40 Proteins in Insect Cells and In Vitro Packaging of Virions and Pseudovirions

  • Protocol
SV40 Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 165))

  • 521 Accesses

The use of insect cells and lytic baculovirus for expression of biologically active mammalian proteins has been the method of choice of many investigators. Although prokaryotic expression systems provide higher yields and are technically simpler to use, obtaining biologically active eukaryotic proteins in these systems can be problematic. Posttranslational modifications such as glycosylation and phosphorylation do not occur in prokaryotic systems owing to the lack of enzymatic machinery. On the other hand, insect cells provide an appropriate environment for posttranslational modifications and lead to proper folding and correct assembly of recombinant proteins (1).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Miller K. L. (1993) Baculoviruses: high-level expression in insect cells. Curr. Opin. Genet. Dev. 3, 97–101.

    Article  CAS  PubMed  Google Scholar 

  2. Summers, M. D. and Smith, G. E. (1988) A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, 2nd ed., Texas Agricultural Experiment Station, College Station, TX.

    Google Scholar 

  3. Luckow, V. A. and Summers, M. D. (1988) Trends in the development of baculovirus expression vectors. Bio/Technology 6, 47–55.

    Article  CAS  Google Scholar 

  4. Luckow, V. A. and Summers, M. D. (1989) High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors. Virology 170, 31–39.

    Article  CAS  PubMed  Google Scholar 

  5. Jeang, K. T., Holmgren-Konig, M., and Khoury, G. (1987) A baculovirus vector can express intron-containing genes. J. Virol. 61, 1761–1764.

    CAS  PubMed  Google Scholar 

  6. Murphy, I. C., Weiner, B., Bikel, I., Piwnica-Worms, H., Bradley, K. M., and Livingston, M. D. (1988) Purification and functional properties of simian virus 40 large and small T antigens overproduced in insect cells. J. Virol. 62, 2951–2959.

    CAS  PubMed  Google Scholar 

  7. Lanford, R. E. (1988) Expression of simian virus 40 T antigen in insect cells using a baculovirus expression vector. Virology 167, 72–81.

    Article  CAS  PubMed  Google Scholar 

  8. Sandalon, Z. and Oppenheim, A. (1997a) Self-assembly and protein-protein interactions between the SV40 capsid proteins produced in insect cells. Virology 237, 414–421.

    Article  CAS  PubMed  Google Scholar 

  9. Sandalon, Z., Dalyot-Herman, N., Oppenheim, A. B., and Oppenheim, A. (1997b) In vitro assembly of SV40 virions and pseudovirions: vector development for gene therapy. Hum. Gene Ther. 8, 843–849.

    Article  CAS  PubMed  Google Scholar 

  10. Carswell, S. and Alwine, J. C. (1986) Simian virus 40 agnoprotein facilitates perinuclear-nuclear localization of VP1, the major capsid protein. J. Virol. 60, 1055–1061.

    CAS  PubMed  Google Scholar 

  11. Resnick, J. and Shenk, T. (1986) Simian virus 40 agnoprotein facilitates normal nuclear location of the major capsid polypeptide and cell-to-cell spread of virus. J. Virol. 60, 1098–1106.

    CAS  PubMed  Google Scholar 

  12. Davis, T. R., Trotter, K. M., Granados, R. R., and Wood, H. A. (1992) Baculovirus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion. Biotechnology 10, 1148–1150.

    Article  CAS  PubMed  Google Scholar 

  13. Davis, T. R., Wickham, T. J., McKenna, K. A., Granados, R. R., Shuler, M. L., and Wood, H. A. (1993) Comparative recombinant protein production of eight insect cell lines. In Vitro Cell Dev. Biol. Anim. 29A, 388–390.

    Article  CAS  PubMed  Google Scholar 

  14. Schreiber, E., Matthias, P., Muller, M. M., and Schaffner, W. (1989) Rapid detection of octamer binding proteins with’ mini-extracts’ prepared from a small number of cells. Nucleic Acids Res. 15, 6419–6436.

    Article  Google Scholar 

  15. Tomer, G. (1993) Expression of SV40 capsid proteins in Escherichia coli. M. Sc. dissertation The Hebrew University, Jerusalem.

    Google Scholar 

  16. Shamri, R. (1998) Production of the SV40 late proteins in Yersinia and E. coli and study of the viral agnoprotein’s function. M. Sc. dissertation. The Hebrew University, Jerusalem.

    Google Scholar 

  17. Gerard, R. D. and Gluzman, Y. (1985) New host cell system for regulated simian virus 40 DNA replication. Mol. Cell. Biol. 5, 3231–3240.

    CAS  PubMed  Google Scholar 

  18. Tooze, J. (1981) DNA Tumor Viruses, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2001 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Sandalon, Z., Oppenheim, A. (2001). Production of SV40 Proteins in Insect Cells and In Vitro Packaging of Virions and Pseudovirions. In: Raptis, L. (eds) SV40 Protocols. Methods in Molecular Biology™, vol 165. Humana Press. https://doi.org/10.1385/1-59259-117-5:119

Download citation

  • DOI: https://doi.org/10.1385/1-59259-117-5:119

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-653-6

  • Online ISBN: 978-1-59259-117-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics